Fig. 5.

Nigericin suppresses vaccinia DNA replication and intermediate/late translation. (A) HeLa cells in 6-well plates were left untreated or were pretreated for 1 h with 2 μM nigericin. Cells were also pretreated with or without 50 μg/mL of cytosine arabinoside (araC) as a positive control for inhibition of vaccinia DNA replication. Cells were then infected with vaccinia virus at an MOI of 1 in the presence or absence each of compound. Cells were collected at 1 and 8 hpi and total DNA was extracted. Real time PCR was performed to quantify the relative abundance of the vaccinia A1 gene. Results are representative of three independent experiments. Error bars represent RQ minimum and RQ maximum values. (B) Cells were treated with nigericin as in (A) and collected at 2 and 8 hpi and the expression of vaccinia virus D12, G8 and H3 was analyzed by Western blotting. Results are representative of three independent experiments.